Antiviral activity of recombinant ankyrin targeted to the capsid domain of HIV-1 Gag polyprotein

作者：Nangola Sawitree; Urvoas Agathe; Valerio Lepiniec Marie; Khamaikawin Wannisa; Sakkhachornphop Supachai; Hong Saw See; Boulanger Pierre^*; Minard Philippe; Tayapiwatana Chatchai

来源：Retrovirology, 2012, 9: 17.

DOI：10.1186/1742-4690-9-17

• HIV-1
• HIV-1 assembly
• Gag polyprotein
• CA domain
• virus assembly inhibitor
• ankyrins
• artificial ankyrin library
• intracellular antiviral agent

Background: Ankyrins are cellular mediators of a number of essential protein-protein interactions. Unlike intrabodies, ankyrins are composed of highly structured repeat modules characterized by disulfide bridge-independent folding. Artificial ankyrin molecules, designed to target viral components, might act as intracellular antiviral agents and contribute to the cellular immunity against viral pathogens such as HIV-1. %26lt;br%26gt;Results: A phage-displayed library of artificial ankyrins was constructed, and screened on a polyprotein made of the fused matrix and capsid domains (MA-CA) of the HIV-1 Gag precursor. An ankyrin with three modules named Ank(GAG)1D4 (16.5 kDa) was isolated. Ank(GAG)1D4 and MA-CA formed a protein complex with a stoichiometry of 1: 1 and a dissociation constant of K-d similar to 1 mu M, and the Ank(GAG)1D4 binding site was mapped to the N-terminal domain of the CA, within residues 1-110. HIV-1 production in SupT1 cells stably expressing Ank(GAG)1D4 in both N-myristoylated and non-N-myristoylated versions was significantly reduced compared to control cells. Ank(GAG)1D4 expression also reduced the production of MLV, a phylogenetically distant retrovirus. The Ank(GAG)1D4-mediated antiviral effect on HIV-1 was found to occur at post-integration steps, but did not involve the Gag precursor processing or cellular trafficking. Our data suggested that the lower HIV-1 progeny yields resulted from the negative interference of Ank(GAG)1D4-CA with the Gag assembly and budding pathway. %26lt;br%26gt;Conclusions: The resistance of Ank(GAG)1D4-expressing cells to HIV-1 suggested that the CA-targeted ankyrin Ank(GAG)1D4 could serve as a protein platform for the design of a novel class of intracellular inhibitors of HIV-1 assembly based on ankyrin-repeat modules.

• 出版日期2012-2-20